Resting posture drives the evolution of agonistic displays in bats.

Agonistic displays are one of the most diverse social behaviors that have important functions in animal's life history. However, their origin and driving factors have largely been unexplored. Here, we evaluated agonistic displays of 71 bat species across 10 families and classified these displays into two categories: (a) boxing displays where a bat attacks its opponent with its wrist and thumb and (b) pushing displays where a bat uses its head or body to hit a rival. We estimated the strength of the phylogenetic signal of the agonistic displays, revealed their origin, and tested the potential evolutionary relationships between agonistic behaviors and body size or resting posture (free hanging vs. contact hanging where the bat is in contact with some surface). We found that agonistic displays were phylogenetically conserved and that boxing displays are the ancestral state. Moreover, we found that bats with a free-hanging resting posture were more likely to exhibit boxing displays than pushing displays. In addition, bats with longer forearms do not have a higher propensity for boxing displays. This study expands our limited knowledge of the evolution of agonistic displays and highlights the importance of resting posture as a driving force in the diversity of agonistic displays.

Population-level lateralization of boxing displays enhances fighting success in male Great Himalayan leaf-nosed bats.

Behavioral lateralization with left- and right-hand use is common in the Animal Kingdom and can be advantageous for social species. The existence of a preferential use of the hands during agonistic interactions has been described for a number of invertebrate and vertebrate species. Bats compose the second largest order of mammals. They not only use their forelimbs for flight but also agonistic interactions. However, whether bat species show a population-level lateralized aggressive display has largely been unexplored. Here, we examine the lateralization of boxing displays during agonistic interactions in male Great Himalayan leaf-nosed bats, Hipposideros armiger, from three different populations. We found a population-level lateralization of boxing displays: Males from all three populations show a preferential use of the left forearm to attack opponents. In addition, left-handed boxers have higher fighting success over right-handed boxers. This study expands our knowledge of the handedness of bats and highlights the role of behavioral lateralization in conflict resolution in nocturnal mammals.

Acoustic signal dominance in the multimodal communication of a nocturnal mammal.

Multimodal communication in animals is common, and is particularly well studied in signals that include both visual and auditory components. Multimodal signals that combine acoustic and olfactory components are less well known. Multimodal communication plays a crucial role in agonistic interactions in many mammals, but relatively little is known about this type of communication in nocturnal mammals. Here, we used male Great Himalayan leaf-nosed bats Hipposideros armiger to investigate multimodal signal function in acoustic and olfactory aggressive displays. We monitored the physiological responses (heart rate [HR]) when H. armiger was presented with 1 of 3 stimuli: territorial calls, forehead gland odors, and bimodal signals (calls + odors). Results showed that H. armiger rapidly increased their HR when exposed to any of the 3 stimuli. However, the duration of elevated HR and magnitude of change in HR increased significantly more when acoustic stimuli were presented alone compared with the presentation of olfactory stimuli alone. In contrast, the duration of elevated HR and magnitude of change in HR were significantly higher with bimodal stimuli than with olfactory stimuli alone, but no significant differences were found between the HR response to acoustic and bimodal stimuli. Our previous work showed that acoustic and chemical signals provided different types of information; here we describe experiments investigating the responses to those signals. These results suggest that olfactory and acoustic signals are non-redundant signal components, and that the acoustic component is the dominant modality in male H. armiger, at least as it related to HR. This study provides the first evidence that acoustic signals dominate over olfactory signals during agonistic interactions in a nocturnal mammal.

Vocal performance reflects individual quality in male Great Himalayan leaf-nosed bats (Hipposideros armiger).

Signals containing parameter trade-offs are likely to be honest indicators of signaler quality because they are difficult to produce. Signals with a trill-rate/bandwidth trade-off have been described for many songbird species, one mouse, and one non-human primate species. However, there were no reports about whether there is a vocal performance trade-off in social calls of bats. This study investigated (1) a possible vocal performance trade-off in territorial calls of male Great Himalayan leaf-nosed bats, Hipposideros armiger, recorded from 9 locations in south China, and (2) the relationships between vocal performance (vocal deviation and consistency) and caller's quality (body mass) to determine whether vocal performance honestly indicates a caller's quality. Vocal deviation measures the deviation of a call relative to an extreme call and vocal consistency measures the spectral consistency across a string of syllables. Our results showed a significant negative correlation between syllable repetition rate and frequency bandwidth, suggesting a vocal performance trade-off similar to the one in songbirds. Further, there was a significant negative relationship between body mass and vocal deviation, but no significant correlation between body mass and vocal consistency. This study provides the first empirical evidence for a vocal performance trade-off of social calls in bats, and the potential for the level of performance to indicate caller quality.

Individuality and function of chemical signals during conflict resolution of a mammal.

Individual recognition via communication signals is a critical component of social behavior, and provides the basis of conflict resolution, territorial behavior, and mate choice. However, the function of chemical signals in mammalian individual recognition and conflict resolution has largely been unexplored despite olfaction being a dominant sensory modality in many mammalian species. Here, we describe behavioral tests designed to evaluate the potential role of forehead gland secretions during conflict related to territorial defense in male Great Himalayan leaf-nosed bats. We used gas chromatography-mass spectrometry to quantify the chemical composition. Our results showed that forehead gland secretions contain 16 categories of compounds, including 84 volatile compounds. The concentrations of compounds and their categories differed significantly among individuals. Moreover, behavioral studies indicated that males can use chemical signals for individual recognition. Contests were staged between males with or without functioning forehead glands. Paired males without functioning forehead glands displayed more physical contact and longer contest duration compared with pairs with functioning glands. Moreover, males with a functioning gland were more likely to win in contests when paired with males without a functioning gland. These findings support a growing amount of evidence that chemical signals play a vital role in conflict resolution in mammals.

Territorial calls of the bat Hipposideros armiger may encode multiple types of information: body mass, dominance rank and individual identity.

In highly vocal species, territorial aggression is often accompanied using vocalizations. These vocalizations can play a critical role in determining the outcome of male-male agonistic interactions. For this, vocalizations of contestants must contain information that is indicative of each competitor's fighting ability as well as its identity, and also contestants must be able to perceive information about the physical attributes, quality and identity of the vocalizer. Here, we used adult male Great Himalayan leaf-nosed bats (Hipposideros armiger) to test whether territorial calls encoded honest information about a caller's physical attributes, quality and individual identity. We did this by exploring the relationships between territorial calls and two potential indices of fighting ability: body mass and dominance rank. Using synchronized audio-video recording, we monitored bat territorial calls and dominance rank of 16 adult male H. armiger in the laboratory. Additionally, habituation-dishabituation playback experiments were performed to test for vocal discrimination. Results showed that body mass was negatively related to minimum frequency and positively related to syllable duration. Dominance score was also negatively related to minimum frequency and positively related to peak frequency. Furthermore, a discriminant function analysis suggested that territorial calls encode an individual signature. Therefore, our data show that males have the ability to utilize this vocal individual signature to discriminate between vocalizing males. In short, territorial calls of male H. armiger contain information about body mass, dominance rank and individual identity, and contestants are probably capable of perceiving this information and may use it to make appropriate decisions during agonistic interactions.

Geographical variation in the echolocation calls of bent-winged bats, Miniopterus fuliginosus.

Evolutionary biologists had a long-standing interest in the evolutionary forces underlying geographical variation in the acoustic signals of animals. However, the evolutionary forces driving acoustic variation are still unclear. In this study, we quantified the geographical variation in the peak frequencies of echolocation calls in eight Miniopterus fuliginosus bat colonies, and assessed the forces that drive acoustic divergence. Our results demonstrated that seven of the colonies had very similar peak frequencies, while only one colony was significantly higher than the others. This similarity in echolocation call frequency among the seven colonies was likely due to frequent dispersal and migration, leading to male-mediated infiltration of nuclear genes. This infiltration enhances gene flow and weakens ecological selection, and also increases interactions in the presence of conspecifics. Significant correlations were not observed between acoustic distances and morphological distances, climatic differences, geographic distances or mtDNA genetic distances. However, variation in acoustic distances was significantly positive correlated with nDNA genetic distance, even after controlling for geographic distance. Interestingly, the relationship between call divergence and genetic distance was no longer significant after excluding the colony with the highest call frequency, which may be due to the minimal genetic distance among the other seven colonies. The highest frequencies of echolocation calls observed in the one colony may be shaped by selection pressure due to loud background noise in the area. Taken together, these results suggest that geographic divergence of echolocation calls may not be subject to genetic drift, but rather, that the strong selective pressure induced by background noise may lead to acoustic and genetic differentiation between JXT and the other colonies.

Endosymbiont CLS-HI plays a role in reproduction and development of Haemaphysalis longicornis.

Coxiella-like endosymbiont (CLS-Hl) is a primary endosymbiont of Haemaphysalis longicornis. CLS-Hl infects tick special tissues and its prevalence is 100% in ovaries and Malpighian tubules. Tetracycline was injected into females, which then fed on rabbits also treated with tetracycline. The densities of CLS-Hl were measured by semi-quantitative PCR. CLS-Hl densities in ovaries and Malpighian tubes of H. longicornis had significant effects on engorged weight, feeding time, number of eggs, oviposition period, and hatching period. These findings suggested that CLS-Hl plays a role in the reproduction and development of H. longicornis.

Microbial diversity of the Tibetan tick Haemaphysalis tibetensis (Acari: Ixodidae).

The Tibetan tick Haemaphysalis tibetensis is widely distributed in the Tibetan Plateau, and is recognized as one of the primary parasites affecting domestic and wild animals. No information is available on its microbial diversity. In the current study, the microbiomes in H. tibetensis were explored using Illumina MiSeq sequencing. Results indicate that a total of 28 phyla and 38 genera were characterized from this tick, and most of the microbes were classified in the phylum Proteobacteria. Female and male H. tibetensis harbored similar composition of the microbes, with dominant bacteria from the genus Coxiella accounting for 87.5 and 66.5% in females and males, respectively. The abundance of bacteria belonging to the genus Rickettsia in females and males was 6.9 and 23.7%, respectively. These findings may help understand the relationship between this tick and its symbiontic microorganisms.

Identification, Distribution and Population Dynamics of Francisella-like Endosymbiont in Haemaphysalis doenitzi (Acari: Ixodidae).

Francisella-like endosymbionts (FLEs) with significant homology to Francisella tularensis (γ-proteobacteria) have been characterized in several tick species, whereas knowledge on their distribution and population dynamics in ticks remains meager. Hence, in the current study, we identified a novel Francisella-like endosymbiont (FLEs-Hd) from the tick Haemaphysalis doenitzi and evaluated the putative functions of this symbiont. Results indicated that FLEs-Hd had 100% infection rate and a perfect vertical transmission in H. doenitzi, and that it is distributed in ovaries, malpighian tubules, salivary glands and midguts of the ticks, suggesting that FLEs-Hd presumably is a crucial symbiont of the host without specific tissue tropism. To further explore the function of the symbiont, the population dynamics of FLEs-Hd at each developmental stage of ticks and in tissues at different reproductive statuses were determined by real-time quantitative polymerase chain reaction (real-time qPCR). Results showed that the high density and regular population dynamics of FLEs-Hd appeared in female ovaries, suggesting that the symbiont may provide necessary nutrients or regulators to ensure normal ovary development of ticks.